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. 2022 May 12;13:2594. doi: 10.1038/s41467-022-30197-8

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Confocal z-projection images of pre- and post-injured aISVs in 3 dpf Tg(fli1a:EYFP-Golgi);(fli1a:mCherry) larva and its subsequent time-lapse images at indicated stages of repair. Images of EYFP-Golgi (green) and mCherry (magenta) are merged. Arrowheads, nuclei. Boxed areas are enlarged on the right. b, c Quantification of front–rear EC polarity in upstream (b) and downstream (c) injured aISVs. Front-rear polarity was analyzed by measuring the angles between the vessel elongation direction and the nucleus-Golgi vector in ECs. Each dot represents an individual cell. Error bars indicate mean ± s.e.m. [the number of ECs over 12 animals: upstream and downstream at pre (n = 15 and 11), post (n = 16 and 7), 25% (n = 15 and 12), 50% (n = 17 and 13), 75% (n = 17 and 14), 100% (n = 16 and 17)]. d Confocal z-projection images of pre- and post-injured aISVs in 3 dpf Tg(fli1a:lifeact-mCherry) larva and its subsequent time-lapse images are shown as in a. e Quantification of F-actin in upstream (red) and downstream (blue) injured aISVs. Relative fluorescence intensity of Lifeact-mCherry in vascular regions within 20 µm from the leading edge is expressed as a percentage of that in pre-injured vessels. Data are means ± s.e.m. [the number of animals: upstream and downstream at pre, post and 50% (n = 5), at 25 and 75% (n = 4), at 100% (n = 3)]. f Confocal z-projection images of pre-injured aISV (left) and injured aISVs at the elapsed time following the injury (hpi) in 3 dpf Tg(fli1a:EGFP-ARPC4);(fli1a:lifeact-mCherry) larva are shown as in a. Images of EGFP-ARPC4 (green) and Lifeact-mCherry (red) are merged. EGFP (top), mCherry (middle), and their merged (bottom) images corresponding to boxed areas are enlarged on the right. g Number of EGFP-ARPC4-labeled Arp2/3 complexes colocalizing with Lifeact-mCherry-marked F-actin in vascular regions within 20 µm from the leading edge at the 25% reparative phase. Each dot represents an individual injured vessel (red and blue, upstream and downstream injured vessels, respectively). Data are means ± s.e.m. (Up and Down, n = 7 and 8 animals). h Confocal z-projection images of pre- and post-injured aISVs and those 3 and 6 h after the beginning of treatment with vehicle or CK-666 in 3 dpf Tg(fli1a:lifeact-mCherry);(fli1a:Myr-EGFP) larva are shown in f. i Amounts of elongation of upstream (red) and downstream (blue) injured aISVs 3 h after the beginning of treatment with vehicle or 200 μM CK-666. Each dot represents an individual injured vessel (red and blue, upstream and downstream injured vessels, respectively). Data are means ± s.e.m. (Up and Down, n = 8 animals). Statistical significance was determined by two-way ANOVA followed by Sidak’s test (e), Welch’s two-sided t test (g), and one-way ANOVA followed by Tukey’s test (i). *p < 0.05. Arrows indicate blood flow direction (a, d, f, h). Source data are provided as a Source data file. Scale bars: 10 µm (a, d, f, h), 5 µm (enlarged images in a, f, h).